Crystalline forms of 4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile and methods of preparing the same

ABSTRACT

This invention is directed to a crystalline 4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile monohydrate having an x-ray diffraction pattern wherein 2θ angles (°) of significant peaks are at about: 9.19, 11.48, 14.32, 19.16, 19.45, 20.46, 21.29, 22.33, 23.96, 24.95, 25.29, 25.84, 26.55, 27.61, and 29.51, and a transition temperature of about 109° C. to about 115° C.

This invention claims the benefit of U.S. Provisional Application No.60/696,381, filed Jul. 1, 2005.

BACKGROUND OF THE INVENTION

1. Field of Invention

This invention is directed to crystalline forms of4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile,methods of preparing these forms and pharmaceutical compositionscontaining them. These compounds are useful in treating cancers,particularly pancreatic and prostate cancer.

2. Related Background Art

3-Cyanoquinoline derivatives have been shown to have anti-tumor activitythat may make them useful as chemoagents in treating various cancers,including pancreatic cancer, melanoma, lymphatic cancer, parotid tumors,Barrett's esophagus, esophageal carcinomas, head and neck tumors,ovarian cancer, breast cancer, epidermoid tumors, cancers of the majororgans, such as kidney, bladder, larynx, stomach, and lung, colonicpolyps and colorectal cancer and prostate cancer.

In the following U.S. patents, 3-cyanoquinoline derivatives aredisclosed and shown to possess anti-tumor activity: U.S. Pat. Nos.6,002,008; 6,432,979; and 6,617,333.

There continues to be a need for forms of4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrilethat are more stable, but still possess a high degree of solubility.

BRIEF SUMMARY OF THE INVENTION

This invention is directed to isolated polymorphs of crystalline4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrileincluding Form I, Form II, Form III, Form IV, Form V and Form VI havingx-ray diffraction patterns as shown in FIG. 1 and FIG. 11. A particularpreferred polymorph is a monohydrate (Form I) having an x-raydiffraction pattern wherein at least one or more, and most preferablyall, of the 2θ angles (°) of significant peaks are at about: 9.19,11.48, 14.32, 19.16, 19.45, 20.46, 21.29, 22.33, 23.96, 24.95, 25.29,25.84, 26.55, 27.61, and 29.51.

Another aspect of this invention is a crystalline4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrilemonohydrate (Form I) having a transition temperature to a liquid ofabout 109° C. to about 115° C.

The invention is also directed to pharmaceutical compositions containinga therapeutically effective amount of crystalline4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile,which has an x-ray diffraction pattern substantially as shown in FIGS. 1and 11 selected from the group consisting of Patterns A, B, C, D, E andF, wherein more than 50% by weight of the crystalline4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrilepresent is one of the selected forms.

This invention is also directed to methods of preparing crystallineforms of4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile,including the monohydrate, alcoholates, and mixtures of both. One methodof preparing4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrilemonohydrate (Form I) comprises the step of treating anhydrous4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile,known herein as Form V, with heated water. Another method of preparing4-[(2,4-Dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrilemonohydrate (Form I) comprises the step of converting other polymorphsof4-[(2,4-Dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile,which have a x-ray diffraction pattern substantially the same as one ofPatterns B, C, D and F, as shown in FIGS. 1 and 11, by treatment withwater. The water can be heated or at a temperature where no heat sourceor cold source is applied, in which case the water would be at roomtemperature.

A BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1. The XRD scans of five different polymorphs of4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile.

FIG. 2. A dynamic vapor sorption isotherm plot of Form I.

FIG. 3. A differential scanning calorimeter plot and atheromogravimetric analysis plot of Form V after suspension in water fortwo months and ten days at room temperature.

FIG. 4. The XRD scans of Form I, Form V and the metastable hydratecreated after Form V is left suspended in water for two months and tendays at room temperature.

FIG. 5. The XRD scan of two batches of Form II.

FIG. 6. The XRD scan of Form III.

FIG. 7. The XRD scan of Form IV.

FIG. 8. The XRD scan of Form V.

FIG. 9. A differential scanning calorimeter plot and atheromogravimetric analysis plot of Form I.

FIG. 10. The XRD scan of five batches of Form I.

FIG. 11. The XRD scans of Form VI (bottom), Form I (top), as a standard,and the form resulting from exposing Form VI to hot water.

DETAILED DESCRIPTION

The present invention is directed to isolated crystalline4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile,which can exist in six different forms, an anhydrous form and fourdifferent hydrated or alcoholated forms. As used herein the term“isolated” means that more than 50% of the crystalline4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrilepresent is one of Forms I, II, III, IV, V and VI and more preferably atleast 70% to 90% of the crystalline4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrilepresent is in one of Forms I, II, III, IV, V and VI.

Form I is a monohydrate and is more stable than the other polymorphforms. It gives substantially Pattern A when scanned by XRD as shown inFIGS. 1 and 10. Form I generally does not lose water when exposed to 0%relative humidity for 10 days, or when heated to 90° C. for over 100hours. Form I is not hygroscopic, as it generally only gains about 0.52%weight when subjected to about 90% relative humidity, as shown by thedynamic vapor sorption plot in FIG. 2. Form I also possesses the highesttransition temperature of all the hydrated forms.

Form II is also a monohydrated form as determined by Karl Fisheranalysis and when scanned by XRD gives substantially Pattern B shown inFIGS. 1 and 5. Form III is a monoisopropyl alcoholate as determined byGLC and NMR. Form IV is likely a hydrated polymorph, though itsstructure is unclear. When scanned by XRD, Forms III and IV, givesubstantially Patterns C and D, as shown in FIGS. 6 and 7, respectively.Form VI is a methanolate form and gives substantially XRD Pattern F, asshown in FIG. 11.

Forms II, III, IV and Form VI can all be transformed into Form I bytreatment with water, e.g. by heating in water. The water may be at roomtemperature. The water may be heated to at least about 90° C., and ashigh as at least about 95° C.

Form V is the anhydrous form, producing substantially Pattern E in FIGS.1 and 8 when scanned by XRD, and has the highest transition temperatureof all the polymorphic forms, i.e., about 148° C. It is also readilyconverted to Form I by treatment with heated water. The water is heatedto at least about 90° C., and more preferably is heated to at leastabout 95° C. In addition, Form V can be converted into a hydrated FormII by treatment with water at room temperature over a period of 2months, as shown by differential scanning calorimetry andthermogravimetric analysis in FIG. 3. This hydrated form is differentfrom either Form I or V, as shown by the XRD scan in FIG. 4. For thisreason Form I is viewed as the more stable form, even though Form V hasthe higher transition temperature.

Shown in Table 1 are 2θ angles from the x-ray diffraction patterns ofForms II, III, IV and V. It should be understood that at least one ofthese peaks must be present in a given form, and preferably at least amajority of the listed peaks will be present for a given form. In a mostpreferred embodiment all the peaks are present for a given form,although one skilled in the art will recognize that whether all thepeaks are observed for a given form may be highly dependent on theconcentration level of the form.

TABLE 1 XRD 2θ Angles for Patterns B (Form II), C (Form III), D (FormIV) and E (Form V). Form II Form III Form IV Form V 2θ 2θ 2θ 2θ 7.626.28 8.58 6.70 9.89 8.64 9.04 10.00 10.49 11.17 11.5 14.45 14.19 11.8112.94 15.27 15.24 12.45 13.42 17.49 16.08 14.11 14.33 19.94 16.76 15.9214.73 20.06 18.46 16.88 16.27 21.65 21.79 18.75 17.46 24.14 23.16 19.4417.83 25.24 24.88 20.70 18.65 26.55 26.36 21.17 19.46 27.40 22.36 20.3828.28 24.88 20.72 28.76 25.42 22.29 26.81 23.02 24.05 26.00

Form I is also more stable, and therefore more desirable, because it canwithstand exposure to various environments for a prolonged period oftime with out degradation. For example, Form I has remainedsubstantially pure and physically unchanged after being exposed to 510foot-candles (˜5490 lux) light for two weeks, 75% humidity at 40° C. for3 months, and 90° C. for 2 weeks, as determined by XRD and HPLC.

Form I has the highest transition temperature of all the hydrated forms.Typically Form I upon heating dehydrates between 95° C. and 100° C. andthen transitions to a liquid in the range of 109° C. to 115° C., but themost likely transition temperature, when substantially pure, is 112° C.The other hydrated forms usually transition to a liquid between 76° C.to 90° C. Table 2 shows the transition temperatures of the variousforms.

TABLE 2 List of different batches in different crystal forms. Solid toLiquid Transition Pattern temperatures and (XRD) CrystallizationComments A refluxed in 95° C. water Transitions at 112° C., monohydrateAcetone/water (3/1) Transitions at 112° C., monohydrate IPA/H20 (1/1)Transitions 109-115° C., monohydrate, low crystallinity Acetone/water(2/1) Transitions at 112° C., monohydrate Acetone/water Transitions at94° C., monohydrate B IPA/H20 (2/1) Transitions at 90° C.,) Converts to“I” in 95° C. water C IPA/H2O (2/1) Transitions at 82° C., 11.5% weightloss by TGA indicates that it is a monoisopropyl alcoholate Converts to“I” in 95° C. water D Acetone/water, (7/2) Transitions at 76° C.Converts to “I” in 90° C. water Acetone/water Transitions at 80° C. EAcetone/water Transitions at 145° C. Converts to “I” in 95° C. water FMethanol Converts to “I” in 95° C. water

The XRD patterns of Forms I, II, III, IV, V or VI can be determined byusing techniques and equipment known to those skilled in the art ofanalytical chemistry and X-ray crystallography. The XRD patterns shownin FIGS. 1, 4, 5, 6, 7, 8 and 11 were produced using X-ray powderdiffraction (Scintag Inc., Cupertino, Calif.), with voltage at 45 kV,current at 40.0 mA, power at 1.80 kW, a scan range (20) of 3° to 40°,scan step size of 0.02°, and a total scan time of 22′38″.

The XRD patterns shown in FIGS. 1, 4, 5, 6, 7, 8, 10 and 11 wereproduced using powder samples. The XRD pattern for each form is uniqueto that form. Each pattern is comprised of a set of diffraction peaks,which can be expressed in 2 theta angles, d-spacing and/or relative peakintensities.

The 2 theta diffraction angles and the corresponding d-spacing valuesaccount for the positions of the peaks found in a XRD pattern. D-spacingvalues are calculated with observed 2 theta angles and copper Ka1wavelength using the Bragg equation. Variations in these numbers canresult from using different diffractometers and also from the method ofsample preparation. However, more variation can be expected for therelative peak intensities. Therefore, identification of the variousforms should be based upon the observed 2 theta angles and thed-spacings, and less importance should be given to the intensities.

Form I has at least one, preferably a majority and most preferably all,of the following characteristic 2 theta angles (°) peaks: 9.19, 9.98,11.48, 14.32, 14.85, 15.64, 19.16, 19.45, 19.71, 20.46, 21.29, 22.33,22.58, 23.96, 24.95, 25.29, 25.84, 26.55, 27.61, 28.42, 29.51, 30.32,31.40, and 32.39.

One skilled in the art would understand that the XRD patterns of FormsI, II, III, IV, V and VI obtained as described herein could containadditional peaks.

The water content of the forms described herein was measured by the KarlFisher method, which is well known to those skilled in the art, using a756 KF Brinkmann Coulometer, with HYDRANAL-WATER Standard 1.00 used asthe standard.

The water content of the hydrated forms described herein, includingFIGS. 3 and 9, was measured using thermogravimetric analysis. A PerkinElmer thermogravimetric analyzer was used for these analyses. Theconditions were a 20 mL/minute nitrogen gas purge, a scan range of 25°C. to 300° C., and a scan rate of 10° C./minute.

The hydroscopicity of the anhydrous Form V and the hydrated forms wasdetermined using dyanamic vapor sorption, including the plot for Form Ishown in FIG. 2. This was performed under the following conditions. RHwas set at 0%, 30%, 52.5%, 75% and 90%, with the sample exposed for 3hours at each RH for two full cycles.

Transition temperatures and heat flow for the various forms, includingthe plot shown in FIGS. 3 and 9, was determined using a Perkin Elmerdifferential scanning calorimeter. The conditions were a 20 mL/minutenitrogen gas purge, a scan range of 25° C. to 300° C., and a scan rateof 10° C./minute.

Pure, crystalline solids have a characteristic transition temperature,the temperature at which point the substance changes state, in thepresent case the solid transitions to a liquid. The transition betweenthe solid and the liquid is so sharp for small samples of a puresubstance that transition temperatures can be measured to 0.1° C.Because it is difficult to heat solids to temperatures above theirtransition temperatures, and because pure solids tend to transition overa very small temperature range, transition temperatures are often usedto help identify compounds. Measurements of the transition temperatureof a solid can also provide information about the purity of thesubstance. Pure, crystalline solids transition over a very narrow rangeof temperatures, whereas mixtures transition over a broad temperaturerange. Mixtures also tend to transition at temperatures below thetransition temperatures of the pure solids.

The crystalline compounds of the present invention may be providedorally, by intralesional, intraperitoneal, intramuscular or intravenousinjection; infusion; liposome-mediated delivery; topical, nasal, anal,vaginal, sublingual, uretheral, transdermal, intrathecal, ocular or oticdelivery. In order to obtain consistency in providing the compound ofthis invention it is preferred that a compound of the invention is inthe form of a unit dose. Suitable unit dose forms include tablets,capsules and powders in sachets or vials. Such unit dose forms maycontain from 0.1 to 300 mg of a compound of the invention and preferablyfrom 2 to 100 mg. Still further preferred unit dosage forms contain 50to 150 mg of a compound of the present invention. The crystallinecompounds of the present invention can be administered orally. Suchcompounds may be administered from 1 to 6 times a day, more usually from1 to 4 times a day. The effective amount will be known to one of skillin the art; it will also be dependent upon the form of the compound. Oneof skill in the art could routinely perform empirical activity tests todetermine the bioactivity of the compound in bioassays and thusdetermine what dosage to administer.

The crystalline compounds of the invention may be formulated withconventional excipients, such as a filler, a disintegrating agent, abinder, a lubricant, a flavoring agent, a color additive, or a carrier.The carrier may be for example a diluent, an aerosol, a topical carrier,an aqueous solution, a nonaqueous solution or a solid carrier. Thecarrier may be a polymer or a toothpaste. A carrier in this inventionencompasses any of the standard pharmaceutically accepted carriers, suchas phosphate buffered saline solution, acetate buffered saline solution,water, emulsions such as an oil/water emulsion or a triglycerideemulsion, various types of wetting agents, tablets, coated tablets andcapsules.

When provided orally or topically, such compounds would be provided to asubject by delivery in different carriers. Typically, such carrierscontain excipients such as starch, milk, sugar, certain types of clay,gelatin, stearic acid, talc, vegetable fats or oils, gums, or glycols.The specific carrier would need to be selected based upon the desiredmethod of delivery, for example, phosphate buffered saline (PBS) couldbe used for intravenous or systemic delivery and vegetable fats, creams,salves, ointments or gels may be used for topical delivery.

The crystalline compounds of the present invention may be deliveredtogether with suitable diluents, preservatives, solubilizers,emulsifiers, adjuvants and/or carriers useful in treatment or preventionof neoplasm. Such compositions are liquids or lyophilized or otherwisedried formulations and include diluents of various buffer content (forexample, Tris-HCl, acetate, phosphate), pH and ionic strength, additivessuch as albumins or gelatin to prevent absorption to surfaces,detergents (for example, TWEEN 20, TWEEN 80, PLURONIC F68, bile acidsalts), solubilizing agents (for example, glycerol, polyethyleneglycerol), anti-oxidants (for example ascorbic acid, sodiummetabisulfate), preservatives (for example, thimerosal, benzyl alcohol,parabens), bulking substances or tonicity modifiers (for example,lactose, mannitol), covalent attachment of polymers such as polyethyleneglycol, complexation with metal ions, or incorporation of the compoundinto or onto particulate preparations of hydrogels or liposomes,micro-emulsions, micelles, unilamellar or multilamellar vesicles,erythrocyte ghosts, or spheroblasts. Such compositions will influencethe physical state, solubility, stability, rate of in vivo release, andrate of in vivo clearance of the compound or composition. The choice ofcompositions will depend on the physical and chemical properties of thecompound capable of treating or preventing a neoplasm.

The crystalline compounds of the present invention may be deliveredlocally via a capsule that allows a sustained release of the compoundover a period of time. Controlled or sustained release compositionsinclude formulation in lipophilic depots (for example, fatty acids,waxes, oils).

One embodiment of the composition of the present invention comprises atherapeutically effective amount of at least one of the crystallineforms of4-[(2,4-dichloro-5-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile,which has an x-ray diffraction pattern substantially as shown in FIGS. 1and 11 selected from the group consisting of Patterns A, B, C, D, E andF, and a pharmaceutical acceptable carrier. In a preferred embodimentmore than 50%, more preferably at least 80%, more preferably greaterthan 90% by weight of the crystalline form is Form I. A more specificembodiment is where this composition comprises either acetic acid or adetergent, or both. More preferably the composition will comprise aceticacid in a weight percentage range of about 0.01% to about 0.1% and thedetergent in a weight percentage range of about 0.5% to about 5.0%. Themost preferred embodiment of this composition is where it is comprisedof about 2.0% by weight detergent and about 0.06% by weight acetic acid.

Another preferred embodiment of the composition of the present inventioncomprises a therapeutically effective amount of at least one of thecrystalline forms of4-[(2,4-dichloro-5-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile,which has an x-ray diffraction pattern substantially as shown in FIGS. 1and 11 selected from the group consisting of Patterns A, B, C, D, E andF, and a pharmaceutical acceptable carrier that is selected from a sugaror polyol or cellulose. In a preferred embodiment more than 50%, morepreferably at least 80%, more preferably greater than 90% of thecrystalline form is Form I, having a x-ray diffraction patternsubstantially the same as Pattern A, as shown in FIG. 1. A morepreferred embodiment is where the sugar or polyol could be mannitol,sorbitol, or xylitol, with mannitol being the most preferred. In a morespecific embodiment the composition would contain both mannitol andcellulose, and more preferably the crystalline forms of4-[(2,4-dichloro-5-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile,the mannitol and the cellulose are individually present in amounts ofabout 20 to about 50 percent by weight. The most preferred embodimentsof the composition of the present invention are shown in Table 3.

TABLE 3 Contents (% w/w) of Compositions Composition No. Ingredients 1 2Crystalline form of 26.32 41.674-[(2,4-dichloro-5-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile A surfactant, such aspoloxamer 3.00 3.00 A disintegrant, such as crospovidone 3.00 3.00Cellulose, micro crystalline 25.00 25.00 Mannitol 42.18 26.83 Alubricant or stabilizer, such as magnesium 0.25 0.25 stearate

4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrileis a weak base with an intrinsic (neutral form) solubility ofapproximately 0.06 μg/mL at pH 8.0. It is partially ionized inde-ionized water where it dissolves to 1.8 μg/mL with a resulting pH of7.2. Solubility is enhanced about 100-fold to 0.1 to 0.4 mg/mL with theaddition of about 2% Tween80, a detergent. Solubility is furtherenhanced to 3-4 mg/mL (˜1000-fold) with the addition of about 0.06%acetic acid (in-situ acetate salt vehicle). The acetate formulation hasthe advantage of eliminating variability in solubility due topolymorphism (see Table 4 below).

4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrilehas poor water solubility and wettability. However, due to the successin solubilizing the compound in the in-situ acetate salt vehicle,micronization is not required for formulation. Furthermore, with thehigh solubility (>200 mg/mL) and better wettability in stomach acid,micronization is also not required for human formulation.

TABLE 4 Solubility Information for4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile.Batch Size (g) 95 50 120 54 18.5 550 Use Rat/Dog MTD(1v) Rat PK(PO) RatMTD (PO) Dog PK(PO) Genotoxicity Dog MTD (PO) Water content % (1¢) 3.683.27 3.24 7.06 3.76 336. Particle Size μm (Malve N/D 50% = 9.87 50% 10.250% 11.7 50% = 10.5 50% = 8.3 90% < 20.2 90% < 30.0 90% < 24.1 90% <29.1 90% < 20.8′ Polymorphism (XRD) Pattern A Pattern A Pattern DPattern D N/D Pattern A Solubility (mg/mL2 Water 0.0018 (pH 7.2) N/D*N/D* N/D* N/D N/D 2% Tween 80/03% MC 0.16 (pH 8.3) 0.20 (pH 8.4) 0.07(pH 7.7) 0.06 (pH 7.8) N/D 0.41 (pH 7.4) 2% Tween80/0.5% MC 4.0 (pH 5.4)N/D 3.5 (pH 5.0) 2.68 (pH 5.2) N/D 4.25 (pH 5.7) 0.06% acetic acid N/D =Not Determined. *Milled batches contained′ a significant amount offines, which could not be removed from solution by centrifugation.Filtration led to erroneously low measurements due to the adsorption ofsolubilized4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrileto the filter material.

The dose provided to a patient will vary depending upon what is beingadministered, the purpose of the administration, the manner ofadministration, and the like. A “therapeutically effective amount” is anamount sufficient to cure or ameliorate symptoms of the disease beingtreated, such as cancer.

The crystalline compounds of this invention may be delivered alone or incombination with other compounds.

4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrilewas prepared as described in Scheme 1.7-(3-chloropropoxy)-4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-3-quinolinecarbonitrile,1, was alkylated with N-methylpiperazine in the presence of sodiumiodide either neat or in a solvent such as ethylene glycol dimethylether. This preparation has been reported in the literature, [Boschelli,D. H., et. al., J. Med. Chem., 44, 3965 (2001)].

DETAILED DESCRIPTION OF THE DRAWINGS

FIG. 1. The XRD scans of five different polymorphs of4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile.The crystalline monohydrate, Form I, is shown as Pattern A, while thecrystalline anhydrous form, Form V, is shown as Pattern E. Patterns B, Cand D are from three other polymorphs of4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile,or mixture of polymorphs, which have not been fully characterized.

FIG. 2. This is a dynamic vapor sorption plot which shows that4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrilemonohydrate, Form I, gained only 0.52% from 0% RH to 90% RH. Therefore,this form must be considered non-hydroscopic.

FIG. 3. This is a differential scanning calorimeter (DSC) plot and atheromogravimetric analysis (TGA) plot of Form V after suspension inwater for two months and ten days at room temperature. The lowtransitioning temperature shown in the DSC and the rapid dehydrationshown in the TGA indicates that after suspension in water for two monthsand ten days indicates that the anhydrate has hydrated to a metastablehydrate crystal form.

FIG. 4. XRD scans of Form I, Form V and Form V after it was suspended inwater for two months and ten days at room temperature, which shows thatthe metastable hydrate which results from this exposure has a differentstructure than either Form I or Form V.

FIG. 5. An XRD scan of two different batches of Form II, which is acrystalline monohydrate of4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile.

FIG. 6. An XRD scan of crystalline4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile in Form III.

FIG. 7. An XRD scan of crystalline4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile in Form IV.

FIG. 8. An XRD scan of the anhydrous crystalline form of4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile, Form V.

FIG. 9. This is a DSC plot and a TGA plot of Form I. The DSC plot showsthat Form I has a transitioning range of about 108° C. to 120° C. TheTGA plot evidences that Form I is a monohydrate since it shows thatthere is approximately a 3.5% loss of weight upon heating to 150° C.

FIG. 10. An XRD scan of five batches of crystalline4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrilemonohydrate in Form I.

FIG. 11. XRD scans of Form VI (bottom), Form I (top), and the formresulting from heating Form VI in water (middle). Thus, it is shown thatForm VI can be converted from into Form I by exposure to water, likeForms II, III, IV and V.

This invention will be more fully described in conjunction with thefollowing specific example, which should not to be construed as limitingthe scope of this invention.

Example 1 Preparation of4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrilemonohydrate, Form I, from Form V

Non-crystalline4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrilewas crystallized from a solution of 50:50 (v/v) acetone and water togive Form V. The resulting crystalline solid was recovered byfiltration, and then suspended in water heated to about 80° C. forapproximately 5 minutes. This mixture was then filtered to yield thetitled compound and having a transitioning range of 109° C. to 115° C.

Example 2 Preparation of4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrilemonohydrate, Form I

4-[(2,4-Dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile(1.8 kg), isopropyl alcohol (12 L), and water (6 L) were added to a 50-Lflask under nitrogen. The mixture was heated to 75° C. and filteredthrough a polypropylene cloth. The mixture was cooled to roomtemperature—crystallization began at about 37° C. The mixture was cooledfurther to 4° C. and then filtered. The cake was washed with 5 L of a50/50 mixture of isopropyl alcohol/water (v/v). The resulting wet cakeand 14 L of water were added to a 50-L flask. The mixture was heated to95° C. and held for 5 hours. The hot mixture was filtered through apolypropylene cloth, filter cake washed with 2.5 L of water, and driedin a forced air oven at 40° C. to give 1.37 kg of4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrileForm I by XRD scans.

Example 3 Preparation of4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile,Form II

4-[(2,4-Dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile(5 g) and 50 mL of a mixture of 2/1 isopropyl alcohol/water (v/v) wereadded to a 125 mL flask, and heated to reflux. The mixture was cooledover night to room temperature and then to 5° C. After filtration, thecake was washed with 15 mL of a 2/1 mixture of isopropyl alcohol/water(v/v) and vacuum dried at 45° C. to give 3.92 g of4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile,Form II by XRD scans.

Example 4 Preparation of4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrilemonoisopropyl alcoholate, Form III

4-[(2,4-Dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile(5 g) and 40 mL of a mixture of 2/1 isopropyl alcohol/water (v/v) wereadded to a 125 flask, and heated to reflux. The mixture was cooled overnight to room temperature and filtered. The cake was washed with 15 mLof a 2/1 mixture of isopropyl alcohol/water (v/v) and vacuum dried at45° C. to give 4.51 g of4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrilemonoisopropyl alcoholate as determined by GLC and NMR and determined tobe Form III by XRD scans.

Example 5 Preparation of4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrilemonoisopropyl alcoholate, Form III

4-[(2,4-Dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile(5 g) and 40 mL of a mixture of 2/1 isopropyl alcohol/water (v/v) wereadded to a 125 flask and heated to 70° C. The hot mixture was filtered;the filtrate was heated to 70° C. to redissolve the material. Themixture was cooled to room temperature. Crystallization began at about45° C. After stirring overnight, the mixture was cooled to 5° C. Thecake was washed with 15 mL of a 2/1 mixture of isopropyl alcohol/water(v/v) and then with 20 mL of water to give 6.58 g of wet4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile.A sample of the wet product (0.30 g) was vacuum dried at 40° C. to give0.19 g of4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrilemonoisopropyl alcoholate as determined by GLC and NMR and determined tobe Form III by XRD scans.

Example 6 Preparation of4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile,Form IV

4-[(2,4-Dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile(5 g) and 30 mL of acetone were added to a 100-mL flask and heated toreflux to give a solution. The hot mixture was filtered to clarify. Thesmall amount of residue on filter paper was washed with 3 ml of acetone.Water (10 ml) was added to the hot filtrate. The mixture was cooled to0-5° C. with an ice-water bath and filtered. The cake was washed with 15ml of a 3/1 mixture of acetone/water (v/v) water and vacuum dried at 45°C. to give 3.48 g of4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile,Form IV by XRD scans.

Example 7 Preparation of4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile,Form VI

4-[(2,4-Dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile(10 g) and 200 mL of methanol were heated to 65° C. and held withstirring for 30 minutes. The hot mixture was cooled to room temperatureand filtered. The material obtained was washed with 80 mL MeOH and driedin vacuo to give 8.31 g of4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrilemethanolate Form VI by XRD scans.

Example 8 Preparation of4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrilemonohydrate, Form I, from Form II

4-[(2,4-Dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile,Form II (1.0 g) was refluxed for 16 hours with 30 ml of water. Themixture was cooled to room temperature and filtered. The cake was washedwith 10 ml of water and vacuum dried at 45° C. to give 0.89 g of4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrilemonohydrate, Form I by XRD scans.

Example 9 Preparation of4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrilemonohydrate, Form I, from Form III

4-[(2,4-Dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile,Form III (2.0 g) was stirred for 24 hours with 40 ml of water at 95° C.The mixture was cooled to room temperature and filtered. The cake waswashed with 10 ml of water and vacuum dried at 45° C. to give 1.83 g of4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrilemonohydrate, Form I by XRD scans.

Example 10 Preparation of4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrilemonohydrate, Form I, from Form III

The wet4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrilemonoisopropyl alcoholate, form III (6.28 g) from example 5, and 40 mL ofwater were added to a 100-mL flask.

The mixture was heated to 95° C. and sampled after 3, 5, and 20 hours.All three samples gave of4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrilemonohydrate, Form I by XRD scans.

Example 11 Preparation of4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrilemonohydrate, Form I, from form IV

4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile,Form IV (1.0 g) was refluxed for 24 hours with 20 ml of water. Themixture was cooled to 35° C. and filtered. The cake was washed with 10ml of water and vacuum dried at 45° C. to give 0.95 g of4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrilemonohydrate, Form I by XRD scans.

Example 12 Preparation of4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrilemonohydrate, Form I, from form VI

4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrilemethanolate, Form VI (1.0 g) was heated with stirring for 5 hours at 95°C. in 12 mL of water. The mixture was cooled to room temperature andfiltered. The cake was washed with 2 mL water and vacuum dried at 45° C.to give 0.9 g of4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrilemonohydrate, Form I by XRD scans.

1-6. (canceled)
 7. A method of preparing4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrilemonohydrate (Form I) comprising the step of converting polymorphs of4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile,which have a x-ray diffraction pattern substantially the same as one ofPatterns B, C, D and F, as shown in FIGS. 1 and 11, by treatment withwater. 8-9. (canceled)
 10. The method of claim 7, wherein the water isat a temperature where no heat source or cold source is applied.
 11. Themethod of claim 10, wherein the water is at room temperature.
 12. Apharmaceutical composition comprising a therapeutically effective amountof a crystalline form of4-[(2,4-dichloro-5-methoxy-7-[3-(4-methyl-3-piperazinyl)propoxy]-3-quinolinecarbonitrile,which has an x-ray diffraction pattern substantially as shown in FIGS. 1and 11 selected from the group consisting of Patterns A, B, C, D, E andF, wherein more than 50% by weight of the crystalline form present isone of the forms selected, and a pharmaceutically acceptable carrier.13. (canceled)
 14. The composition of claim 12, further comprisingacetic acid.
 15. The composition of claim 12, further comprising adetergent.
 16. The composition of claim 12, further comprising about0.5% to about 5.0% by weight of a detergent and about 0.01% to about0.1% by weight acetic acid.
 17. The composition of claim 12, furthercomprising about 2.0% by weight detergent and about 0.06% by weightacetic acid.
 18. The composition of claim 12, wherein thepharmaceutically acceptable carrier is selected from a sugar, a polyoland cellulose, or a combination thereof.
 19. The composition of claim18, wherein the pharmaceutically acceptable carrier is a polyol andcellulose.
 20. The composition of claim 19, wherein the polyol isselected from mannitol, sorbitol and xylitol.
 21. The composition ofclaim 20, wherein the polyol is mannitol.
 22. The composition of claim21, wherein the crystalline form of4-[(2,4-dichloro-5-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile,the mannitol and the cellulose are individually present in amounts ofabout 20 to about 50 percent by weight.
 23. An isolated crystalline formof4-[(2,4-dichloro-5-methoxy-7-[3-(4-methyl-3-piperazinyl)propoxy]-3-quinolinecarbonitrilemonohydrate having an x-ray diffraction pattern substantially as shownin FIG.
 5. 24-28. (canceled)